LABORATORY


Carefull processing of your sample material performed by experienced hands in optimal facilities


To prepare your material for deep-sequencing, three essential steps are required in the laboratory. At omiics, we have the facilities and experience to conduct all three for you.  

RNA EXTRACTION


Isolation of RNA from cells, tissues or biofluids



LIBRARY PREPARATION


Prepare RNA for sequencing choosing the optimal kit



QUALITY CONTROL


Test the library quality before sequencing

Today there is a wide selection of library preparation kits available and which one is the best choice depends on both the input quality and quantity, as well as, the type of end-analysis needed.


Make it easy for yourself - Let omiics choose the optimal reagents for your setup. 

Library preparation

RNA EXTRACTION

To perform sequencing of your sample, we initially isolate RNA.


The optimal procedure depends on whether you are interested in sequencing total/long RNA (mRNA, lncRNA, circRNA) or small RNAs (miRNA, tRNA), and the sample material.

Bioliquids - liquid biopsies


Even cell-free liquids may contain RNA that have been secreted from cells in the local tissue, or transfered from other parts of the body by blood or lymph. 


A shift in RNA patterns can be an early sign of changes in the body and as such has a strong potential as biomarker, both by invasive (CSF, plasma, bile) and  non-invasive testing (saliva, urine, sweat). 



    Sample material


    omiics has experience with a vast array of biological sample material. Examples include muscle and brain tissue, mammalian cells, exosomes, blood, CSF, IVF media, bile, saliva and food powders. 




    Sample quality

    We specialize in low-input samples (e.g. cell-free material) and have successfully sequenced samples using input as low as 250 pg RNA. For standard setups, we recommend input of 500 ng (total RNA) to 1 μg (small RNA) for sequencing.  


    Sample quantity

    RNA is a highly unstable molecule. It is crucial to handle samples in an RNAse-free environment and keep samples below 4 °C at all times. As a standard, RNA should have a RIN value above 7 for optimal sequencing. 

    When optimal

    is not possible..

    Unique material comes in limited amounts and may for different reasons be of low quality. Also, sometimes samples have been isolated by measures essential to adress novel questions, yet damaging the RNA. It may hinder some types of analysis, but far from all.

    In omiics, we have extensive experience with getting information from even low quality samples. 


    Lets talk about what we can extract from your preciouses samples

    Deplete the many or enrich the few?


    Ribosomal RNA (rRNA) comprises a significant proportion (~90% or more) of all the RNA in total RNA samples. While the rRNA will not directly cause harm to the result, removing rRNA or enrichment of mRNA prior to generating libraries provides benefits by lowering sequencing costs and improving mapping statistics.

    We highly recommend using rRNA removal as this provides data on both coding and noncoding RNAs. While historically polyA enrichment has been cheaper, the current prices are comparable,

    - so why settle for less infomation? 

    omiics recommend

    Unique molecular identifyer (UMI)


    A speciality of omiics is sequencing ultra-low input samples down to as low as 250pg RNA.

    At ultra-low RNA concentration, the sample becomes more sensitive to PCR-associated bias. By including an additisonal molecular tag during library preparation, the unique molecular identifyer, UMI, enables you to get high-resolution reads, to facilitate accurate detection of true variants.

     


    Quality control

    For each step of the process in the laboratory we have strict quality control. If for some reason, a sample fail to pass our standards, we will contact you and discuss the options.    

    RNA quality 


    The quality and concentration of extracted RNA is analyzed by Qubit and Bioanalyser 2100.  


    Based on concentration and the RIN value it is decided if the material is appropriate for the contracted sequencing design.

    Library quality


    Library quality is determined using the Bioanalyzer 2100, and accurate concentration of sequencable library is estimated by qPCR.

     

    Once passed, the next step is sequencing